The Analysis Core will provide expertise, personnel and equipment for molecular analysis methods relevant to gene therapy and the response of cells and tissues to gene transfer. The Analysis Core is located in the newly renovated Belfer Gene Therapy Core Facility and consists of laboratory space and an equipment room containing the Sequence Detection System 7700 (TaqMan) realtime quantitative PCR system and BioMek FX sample preparation robot. The Core will make available to the investigators of the Program Project several advanced molecular techniques relevant to gene therapy. These include: automated DNA extraction and purification; TaqMan quantitative PCR for DNA quantitation and quantitative RT-PCR for assessment of changes in gene expression levels. Also available, although not specifically part of the proposed studies in the Program Project, are high throughput genotyping, gene expression microarrays (Affymetrix GeneChip), and analysis of genomic and microarray expression data. All four projects will make equal use of the Analysis Core, primarily in the area of quantitative PCR for quantitation of vector DNA in cells and tissues and in quantitative RTPCR for the absolute and relative quantitation of gene expression levels. The Director of the Analysis Core, Neil Hackett, has spent considerable time optimizing quantitative PCR (TaqMan) methods to assess gene transfer and demonstrated the potential power and high throughput of the method when properly executed. Due to the complexity and expense of molecular techniques and the advanced equipment required, it is a challenge for any one group to acquire all of the sophisticated methodologies and efficiency derived from high throughput that are available through the Analysis Core.